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Journal of Animal Research: v.3 n.1 p.99-102. June, 2013
Antibiotic resistance pattern among different Listeria species isolated from mutton and chevon Smrati Gupta1 and Varsha Sharma2*
1Veterinary Assistant Surgeon, Behind Sardar Patel School,Gharaula Mohalla,Distt. Shahdol (M.P.), INDIA2Department of Veterinary Microbiology, College of Veterinary Science & AnimalHusbandry, Mhow (M.P.), INDIA
*Corresponding Author: V. Sharma; Email: drvsharma04@yahoo.co.in
ABSTRACT
In the present study, Listeria were isolated and confirmed from 50 mutton and 50chevon samples and their antibiotic resistance pattern was studied against 18commonly used antibiotics. Out of 100 samples 4 Listeria isolates r evealed )resistance against cephotaxime and cloxacillin. Similarily, resistance was observedto cephotaxime, chloramphenicol, cloxacillin and oxytetracycline among two L. welshimeri isolates isolates) whereas L. innocua isolates were resistant resistantto cephotaxime, cefoperazone, cloxacillin, oxytetracycline and gentamicin. Potential transmission of multidrug-resistant from food animals to humans is aserious concern in zoonotic pathogens like ListeriaKeywords: Mutton , Chevon , Listeria, multidrug-resistant zoonotic pathogens
A peculiar property of Listeria that affects its food-borne transmission is the abilityto multiply and grow in temperatures ranging from temperature of a refrigerator to37°C (99°F), the body’s internal temperature (Southwick and Purich, 2007; Winteret al.,2004). The resistance to antibiotics has been advocated as a major cause oftreatment failure and anti microbial sensitivity testing has been adopted to improveefficacy. Animals are reservoir of gram negative bacteria harbouring antimicrobialresistance. The continuous introduction of new antibiotics and their widespread useand never changing pattern of drug resistance emphasis the importance of the in-vitro testing for antibiotic susceptibility profile. MATERIALS AND METHODS
All the confirmed Listeria isolates which were recovered from 50 Mutton and 50Chevon samples comprising muscle and viscera were subjected for antibioticsensitivity. The in vitro antibiotic sensitivity tests of the isolates were conducted
with minor modifications (Bauer et al., 1966) as follows. In brief, a loopful ofthe growth from slant was inoculated in BHI broth and incubated at 37oC for 3 to5 hrs. The opacity of broth tube was matched with McFarland’s tube No. 5 (1.5× 106 organisms/ml). A sterile cotton swab was dipped into the broth culture,excess of the bacterial suspension was removed by pressing and rotating theswab against the inner walls of the test tube. Streak the entire agar surface of theplate with the swab three times, turning the plate at 60o angle between eachstreaking. The surface of pre-incubated and sterile Muller-Hinton agar (Hi MediaLtd., Mumbai) petri plate was kept at room temperature for 30 min to allow theinoculum to be adsorbed on the surface. Antibiotic sensitivity disc (Table 1; HiMedia Ltd, Mumbai) were placed with the help of flamed forceps on the plates atequal distance and sufficiently separated from each other. The plates were incubatedovernight at 37°C. Antibiotic sensitivity disc were used i.e. amoxycillin, amoxyclav,ampicillin, azithromycin, cefoperazone, cephalothin, cephotaxime,chloramphenicol, chlortetracycline, ciprofloxacin, cloxacillin, cotrimoxazole,enrofloxacin, erythromycin, gentamicin, oxytetracycline, norfloxacin andtetracycline . Diameters of the clear zone of inhibition were measured and theinterpretation of the results was made in accordance with the instructions suppliedby the manufacturer. Table 1: Zone size interpretative chart for in vitro antibiotic sensitivity. RESULTS AND DISCUSSION
All the 4 isolates of Listeria were tested for in vitro sensitivity towards 18antibacterial drugs. Sensitivity of individual isolate to various drugs was interpreted
Journal of Animal Research: v.3 n.1 p.99-102. June, 2013
Antibiotic resistance pattern among different Listeria species isolated
according to the manufacturer’s instructions. In this study Listeria isolates werefound variably sensitive and resistance to the antibiotics tested. In general, mostof isolates were sensitive to chlortetracycline (100%), higher percent of isolateswere sensitive amoxycillin (75%), enrofloxacin (75%), amoxyclav (75%),cephalothin (75%), ciprofloxacin (75%), tetracycline (75%) and norfloxacin (75%),While moderately high percent of isolates were sensitive to oxytetracycline (50%),co-trimoxazole (50%), chloramphenicol (50%), cefoperazone (50%), azithromycin(50%), ampicillin (50%) and lesser per cent of isolates were observed sensitive toerythromycin (25%), gentamicin (25%) while isolates were observed resistantagainst cephotaxime (0%) and cloxacillin (0%).Out of 4 Listeria isolates two L. welshimeri isolates sensitive to the amoxicillin (100%), chlortetracycline (100%),cefoperazone (100%) While moderately high percent of isolates were sensitive toamoxyclav (50%), ampicillin (50%), azithromycin (50%), ciprofloxacin (50%),cephalothin (50%), co-trimoxazole (50%), norfloxacin(50%), enrofloxacin (50%),gentamicin (50%), tetracycline (50%) and erythromycin (50%) and resistence tocephotaxime (0%), chloramphenicol (0%), cloxacillin (0%) and oxytetracycline(0%). L. innocua isolates sensitive to the amoxyclav (100%), chloramphenicol(100%), ciprofloxacin (100%), cephalothin (100%), norfloxacin (100%),chlortetracycline (100%), enrofloxacin (100%), tetracycline (100%) andoxytetracycline (100%), while moderately sensitive to ampicillin (50%), co-trimoxazole (50%), azithromycin (50%) and amoxicillin (50%), while cent percentresitance to cephotaxime, cefoperazone, cloxacillin, oxytetracycline and gentamicin.
The present findings were in partial agreement with that of Yatiraj (2008) whoreported sensitive to chloramphenicol (96.29%), amoxycillin (87.03%), enrofloxacin(83.34%), amoxyclav (77.78%), tetracycline (68.51%), ampicillin (57.40),oxytetracycline (48.15%), streptomycin (40.74%), co-trimoxazole (33.33%),erythromycin (27.78%) and gentamicin (14.81%).
Kumar et al. (2005) reported multidrug resistant Listeria. Antibiotic sensitivity of 14isolates revealed maximum resistance against cloxacilin (100%) followed by vancomycin(92.85%), amoxycillin, cephalothin and amoxyclav (85.71% each), erythromycin(78.57%), clindamycin and co-trimoxazole (70% each). The maximum sensitivity wasobserved with ciprofloxacin and tetracycline (66.66 % each). Yadav (2008) reportedListeria isolates to be variably resistant to the antibiotics. REFERENCES
Bauer, A.W., Kirby, W.M. Shernis M.J.C. and Turek M. 1966. Antibiotic susceptibility testing
by standard single disc diffusion method. American Journal of Clinical Pharmacology, 45: 493-496.
Kumar, R., Agarwal, R.K., Bhilegaongar, K.N, Garg, A.P., Tyagi, K. and Puroshottam, K.
2005. Occurrence of multidrug resistant Listeria Spp. in meats and fish. Journal of Veterinary Public Health.3: 13-18.
Southwick, F.S. and Purich, D.L. 2007. University of Florida Medical School. http://
Journal of Animal Research: v.3 n.1 p.99-102. June, 2013
www.med.ufl.edu/ biochem/DLPURICH/ morelist html.
Winter, P., Schilcher,F., Bagò, Z., Schoder, D., Egerbacher, M., Baumgartner, W. and Wagner, W.
2004. Clinical and Histopathological Aspects of Naturally Occurring Mastitis Caused by Listeria monocytogenes in Cattle and Ewes. Journal of Veterinary Medicine,51: 176–179.
Yadav, M.M. 2008. PhD Thesis, Anand Agricultural University, Anand, Gujarat. Yatiraj, S.N. 2008. M.V.Sc. and A.H. Thesis, Jawaharlal Nehru Krishi Vishwa Vidyalaya,
Journal of Animal Research: v.3 n.1 p.99-102. June, 2013
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